Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 720416 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- KChIP2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Bovine, Cat.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of KCNIP2 was achieved by transfecting SH-SY5Y cells with KCNIP2 specific siRNA (Silencer® select Product # s26930 + s26931). Western blot analysis (Fig a) was performed using Membrane enriched extracts from the KCNIP2 knockdown cells (Lane 2) and non-specific scrambled siRNA transfected cells (Lane 1). The blots were probed with Anti-KCNIP2 Rabbit Polyclonal Antibody (Product # 720416, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.2 µg/mL, 1:5000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to KCNIP2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), Jurkat (Lane 2), MOLT-4 (Lane 3) and HeLa (Lane 4). The blots were probed with Anti-KCNIP2 Rabbit Polyclonal Antibody (Product # 720416, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.2 µg/mL, 1:5000 dilution). A 31 kDa band corresponding to KCNIP2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, SH-SY5Y cells were fixed for detection of endogenous KCNIP2 using Anti- KCNIP2 Rabbit Polyclonal Antibody (Product # 720416, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of KCNIP2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating Membrane localization of KCNIP2 . Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
